PROJECTSUMMARY Inthisrenewalapplication,theScientificCore(CoreB)willcontinuetobuilduponthestrengthsofthe HumanImmuneMonitoringCenter(HIMC)andtheassaysdevelopedduringtheinitialgrantperiod.The HIMCisanidealenvironmentforstandardization,technologydevelopment,anddataanalysis,andhasbuilt anarrayofhigh-throughputandhigh-contentassaysthatareunparalleledinasinglecenterandinstitution. Overtheinitialfundingperiod,theScientificCoreaddedtothismenuoftechnologieswithuniqueCyTOF panelsandcombinedTCRseq/geneexpressionassaysatthesingle-celllevel.Overtheproposednext fundingperiod,CoreBwillcontinuetoprovidestandardizedimmunologicalassaysacrossstudieswithinthe U19project,focusingonsingle-cellassaysofallergen-responsivecellsandglobalimmuneprofiling.Itwill alsocontinuetodevelopnovelapproachesandworkwiththeindividualprojectstominetheimmunological datagenerated.Thespecificaimsare: 1.Toprovidestandardizedimmunemonitoringassaysacrossprojects.Thesewillincludedetailed immunophenotypingofwholebloodandPBMCsbymasscytometry(CyTOF),includingabasophil-centric CyTOFpaneldevelopedinthelabofSeanBendall(Co-InvestigatorofCoreB).TheCorewillalsooffer62- plexLuminexcytokinearraystoassessserumandcellsupernatantsforTh1/Th2/Th17cytokinesandsingle- cellTCRseq+geneexpressionforallergen-specificTcells.Finally,theCorewillperformMultiplexedIon BeamImaging(MIBI)ofesophageal,gastricandduodenalbiopsyspecimens,inthelabofRobertAngelo (Co-InvestigatorofCoreB).Byusingthesamestandardizedassaysacrossallcohorts,wewillgreatly increasethepowertotesthypothesesrelatedtoanyofthereadoutsoftheseassays. 2.Todevelopnovelimmunemonitoringassayswithindividualprojects.TheCyTOF,MIBI,andTCRseq panelswillcontinuetobedevelopedbasedonnewknowledgegeneratedwithintheprojects,aswellaskey observationsmadeintheallergyfieldgenerally.Wealsoplantoofferanewimmunoassayplatform, SomaScanfromSomaLogic,for1300solubleproteinanalytes,asanadditionaldiscoverytool. 3.Tofacilitateminingofimmunemonitoringdata.UsingStanfordDataMiner(SDM),theCorewillwork withtheDataManagementandAnalysisCoreCtominedatausingbioinformaticstools.SDMwillprovidea structuredframeworkforextractinglaboratorydatalinkedtoclinicalanddemographicvariables,facilitating dataminingandreducingthepossibilityoferrors.